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WO 2004/096978 



PCT/IL2004/000181 




SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCT/IL2004/000181 



2/17 

I Kpnl (1652) 




Fig. 1b 

SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCT/IL2004/000181 



3/17 



1 2 3 4 5 




Fig. 2 

Transformed cells express rGCD. 1 gram calli tissue was 
homogenized and 15 microgram of soluble cell extract were 
run on SDS-PAGE. Expression of rGCD in selected transformed 
calli was tested by western blot analysis with specific anti hGCD 
antibodies. 1: standard cerezyme, 2: untransformed callus extract, 
3-5: various selected transformed calli extracts . 



SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCT/IL2004/000181 



4/17 




SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCT/IL2004/000181 




SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCT/IL2004/000181 



6/17 




SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



7/17 



PCT7IL2004/000181 



4.0 




0 2 4 6 8 1012 14161820 22 24 26 28 3032 34 36 38 40 4244 46 48 50 



tube number 

Fig. 3e 




MW kDa 


A2 17 18 19 20 21 22 23 


^ 175 




83 




■:■ 62 ! 




,47.5 




32.5 




25 


Fig. 3f 



SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 PCT/IL2004/000181 



8/17 




SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCTAL2004/000181 




SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 PCT/IL2004/000181 

10/17 



0.30- 
0.25- 
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Affect of Mannan on rGCD uptake 



o r-GCD 

o r-GCD-mannan 



10 
ug GCD 



20 



40 



Fig. 5a 



SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCT/IL2004/000181 



11/17 



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SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCT/IL2004/000181 



12/17 



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SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCT/IL2004/000181 



13/17 




Major glycan structure 
from CHO cells 



Glycan 

remodeling with 
glycosidases 




Major remodeled glycan 
structure on Cerezyme 




Fig. 6 



Major glycan structure 
from carrot cells: 

Mannose terminal glycan 



NeuAc © 

Gal <3> 

Man © 

Xyl 

Fuc A 
GlcNac |T~] 



a) Ma3 — Mb4GNb4 — GN b) Ma3 — Mb4GNb4GN 
Ma6-| I Ma6-+ 

Xb2-+ Fa3-+ 



c) Ma3 — Mb4GNb4 — GN 
Ma6-+ I 
Fa3-+ 



Fig. 7 



SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCT/BL2004/000181 



14/17 




Theoretical monoisotopic mass for [M+Na] molecular ion = 1171.5 




Theoretical monoisotopic mass for [M+Na] molecular ion = 1331 .6 




Theoretical monoisotopic mass for [M+Na] molecular ion = 1345.6 




Theoretical monoisotopic mass for [M+Na] molecular ion = 1505.7 



Fig. 8a 



SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCT/IL2004/000181 



15/17 




Theoretical monoisotopic mass for [M+Na] + molecular ion = 1579.8 



v1 3/6, 




Theoretical monoisotopic mass for [M+Na] molecular ion = 1709.7 




Theoretical monoisotopic mass for [M+Na] + molecular ion = 1750.9 



O 1 - 2 




Theoretical monoisotopic mass for [M+Na] + molecular ion = 1783.9 

Fig. 8b 



SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCT/IL2004/000181 



16/17 



O- 2 




Theoretical monoisotopic mass for [M+Na] molecular ion = 1989.0 
11 2 

p. 

1 4 




Theoretical monoisotopic mass for [M+Na] + molecular ion = 1997.0 




Theoretical monoisotopic mass for [M+Naf molecular ion = 2130.0 



O- 2 



LO 




Theoretical monoisotopic mass for [M+Na] + molecular ion = 2193.1 

Fig. 8c 



SUBSTITUTE SHEET (RULE 26) 



WO 2004/096978 



PCT/IL2004/000181 



17/17 




Theoretical monoisotopic mass for [M+Na] molecular ion = 2375.2 




Theoretical monoisotopic mass for [M+Na] molecular ion = 2375.2 



Fig. 8d 



Key: Fucose 

® Galactose 

[_J N-Acetylglucosamine 
Mannose 

O Xylose 



SUBSTITUTE SHEET (RULE 26)